scholarly journals Large-scale label-free single-cell analysis of paramylon in Euglena gracilis by high-throughput broadband Raman flow cytometry

2020 ◽  
Vol 11 (4) ◽  
pp. 1752 ◽  
Author(s):  
Kotaro Hiramatsu ◽  
Koji Yamada ◽  
Matthew Lindley ◽  
Kengo Suzuki ◽  
Keisuke Goda
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Euglena Gracilis ◽  
Large Scale ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Label Free

scholarly journals High-throughput label-free molecular fingerprinting flow cytometry

2019 ◽  
Vol 5 (1) ◽  
pp. eaau0241 ◽  
Author(s):  
Kotaro Hiramatsu ◽  
Takuro Ideguchi ◽  
Yusuke Yonamine ◽  
SangWook Lee ◽  
Yizhi Luo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Fluorescent Labeling ◽  
Live Cells ◽  
Molecular Fingerprinting ◽  
Label Free ◽  
Indispensable Tool

Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm−1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics ofHaematococcus lacustris.

Single-Cell Analysis of Morphological and Metabolic Heterogeneity in Euglena gracilis by Fluorescence-Imaging Flow Cytometry

2018 ◽  
Vol 90 (19) ◽  
pp. 11280-11289 ◽  
Author(s):  
Hector E. Muñoz ◽  
Ming Li ◽  
Carson T. Riche ◽  
Nao Nitta ◽  
Eric Diebold ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Fluorescence Imaging ◽  
Euglena Gracilis ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Imaging Flow Cytometry ◽  
Metabolic Heterogeneity

scholarly journals Droplet flow cytometry for single-cell analysis

RSC Advances ◽  
2021 ◽  
Vol 11 (34) ◽  
pp. 20944-20960
Author(s):  
Ming Li ◽  
Hangrui Liu ◽  
Siyuan Zhuang ◽  
Keisuke Goda
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Droplet Flow ◽  
Recent Advances

This work reviews recent advances in the integration of emulsion microdroplets and flow cytometry technologies, so-called droplet flow cytometry (DFC), for high-throughput single-cell analysis.

scholarly journals Noninvasive High-Throughput Single-Cell Analysis of HIV Protease Activity Using Ratiometric Flow Cytometry

Sensors ◽  
2013 ◽  
Vol 13 (12) ◽  
pp. 16330-16346 ◽  
Author(s):  
Rok Gaber ◽  
Andreja Majerle ◽  
Roman Jerala ◽  
Mojca Benčina
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Protease Activity ◽  
Single Cell Analysis ◽  
Hiv Protease ◽  
Cell Analysis

scholarly journals Optimized double emulsion flow cytometry with high-throughput single droplet isolation

2019 ◽  
Author(s):  
Kara K. Brower ◽  
Catherine Carswell-Crumpton ◽  
Sandy Klemm ◽  
Bianca Cruz ◽  
Gaeun Kim ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Double Emulsion ◽  
Droplet Microfluidics ◽  
Cell Analysis ◽  
Single Droplet ◽  
Emulsion Droplets ◽  
Droplet Sorting

Droplet microfluidics has made large impacts in diverse areas such as enzyme evolution, chemical product screening, polymer engineering, and single-cell analysis. However, while droplet reactions have become increasingly sophisticated, phenotyping droplets by a fluorescent signal and sorting them to isolate variants-of-interest remains a field-wide bottleneck. Here, we present an optimized double emulsion workflow, sdDE-FACS, that enables high-throughput phenotyping, selection, and sorting of droplets using standard flow cytometers. Using a 130 μm nozzle, we demonstrate robust post-sort recovery of intact droplets, with little to no shear-induced droplet breakage, at high sort frequency (12-14 kHz) across two industry-standard FACS instruments. We report the first quantitative plate statistics for double emulsion droplet isolation and demonstrate single droplet recovery with >70% efficiency. In addition, we establish complete downstream recovery of nucleic acids from single, sorted double emulsion droplets, an advance in droplet sorting comparable with the capabilities of single-cell FACS. This work resolves several hurdles in the field of high-throughput droplet analysis and paves the way for a variety of new droplet assays, including rare variant isolation and multiparameter single-cell analysis, marrying the full power of flow cytometry with droplet microfluidics.

scholarly journals Towards high-throughput microfluidic Raman-activated cell sorting

The Analyst ◽  
2015 ◽  
Vol 140 (18) ◽  
pp. 6163-6174 ◽  
Author(s):  
Qiang Zhang ◽  
Peiran Zhang ◽  
Honglei Gou ◽  
Chunbo Mou ◽  
Wei E. Huang ◽  
...  
Keyword(s):  
Single Cell ◽  
High Throughput ◽  
Cell Sorting ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Label Free ◽  
Non Invasive

Raman-activated cell sorting (RACS) is a promising single-cell analysis technology that is able to identify and isolate individual cells of targeted type, state or environment from an isogenic population or complex consortium of cells, in a label-free and non-invasive manner.

scholarly journals Noninvasive High-Throughput Single-Cell Analysis of the Intracellular pH of Saccharomyces cerevisiae by Ratiometric Flow Cytometry

2013 ◽  
Vol 79 (23) ◽  
pp. 7179-7187 ◽  
Author(s):  
Mari Valkonen ◽  
Dominik Mojzita ◽  
Merja Penttilä ◽  
Mojca Benčina
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Flow Cytometry ◽  
Single Cell ◽  
Intracellular Ph ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Cell Physiology ◽  
Complex Data ◽  
Cell Analysis ◽  
Ph Homeostasis

ABSTRACTThe ability of cells to maintain pH homeostasis in response to environmental changes has elicited interest in basic and applied research and has prompted the development of methods for intracellular pH measurements. Many traditional methods provide information at population level and thus the average values of the studied cell physiological phenomena, excluding the fact that cell cultures are very heterogeneous. Single-cell analysis, on the other hand, offers more detailed insight into population variability, thereby facilitating a considerably deeper understanding of cell physiology. Although microscopy methods can address this issue, they suffer from limitations in terms of the small number of individual cells that can be studied and complicated image processing. We developed a noninvasive high-throughput method that employs flow cytometry to analyze large populations of cells that express pHluorin, a genetically encoded ratiometric fluorescent probe that is sensitive to pH. The method described here enables measurement of the intracellular pH of single cells with high sensitivity and speed, which is a clear improvement compared to previously published methods that either require pretreatment of the cells, measure cell populations, or require complex data analysis. The ratios of fluorescence intensities, which correlate to the intracellular pH, are independent of the expression levels of the pH probe, making the use of transiently or extrachromosomally expressed probes possible. We conducted an experiment on the kinetics of the pH homeostasis ofSaccharomyces cerevisiaecultures grown to a stationary phase after ethanol or glucose addition and after exposure to weak acid stress and glucose pulse. Minor populations with pH homeostasis behaving differently upon treatments were identified.

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